Title of Invention

"A PROCESS FOR THE PREPARATION OF EXTRACT CONTAING 24 TO 33% ANTIOXIDANTS FROM DILLENIA INDICA

Abstract

1. A process for the preparation of extract conting 24 to 33% antioxidants from Dillenia indica, which comprises: i) powdering the fruit rinds of Dillenia indica using mixer grinder to mesh size of 40-80, ii) extracting the above said material with hexane in a Soxhlet extractor at a temperature of 55-60 °C for a period of 6-10 h., iii) distilling the above said extract to recover up to 90% the solvent, iv) extracting the spent obtained from step (ii) with an alcoholic solvent in a Soxhlet extractor at a temperature of 55-60 °C for a period of 6-10 hrs., v) filtering the above said extract to obtain the particle free extract, vi) distilling the above said particle free extract at a temperature of 55 - 60 °C to recover about 90% solvent, vii) drying the above concentrated extract using vacuum oven at a temperature of 40-50 °C, at a pressure of 10-25 mm of mercury under vacuum to obtain .the extract conting 24 to 33% antioxidants

Full Text

The present invention relates to a process for the preparation of antioxidants from Dillenia indica. The notion that deleterious effects of oxidative metabolism can be ameliorated through a diet rich in antioxidants has gained credibility and the term " functional foods" (Strain, J.J.; Benzie, I.F.F. Antioxidant nutrients. In: Sadler, M.J. and Saltmarsh, M. Eds. Functional Foods: The Consumer, the Products and the Evidence. Cambridge, U.K., Royal Society of Chemistry, p. 74) has been applied to foods with these nutrients. It is important to note that most of the investigations regarding the inhibiting effects of food components on the oxidative damages of biological materials have been devoted to the foods of plant origin. Phenolics are the dominant antioxidants found in the natural products of plant sources (Prior, R.L. and Cao, G. Variability in Dietary antioxidant related natural product supplements: The need for methods of standardization, Journal of the American Nutraceutical Association, 2, 46-56, 1999). Antioxidants are the compounds which when added to food products, specially to lipids and lipid containing foods, can increase the shelf life by retarding the process of lipid peroxidation, which is one of the major reasons for deterioration of food products during the processing and storage. Synthetic antioxidants such as butylated hydroxyanisole (BHA) and butylatedhydroxy toluene (BHT), propylgallate (PG) and tert-butylhydroquinone (TBHQ) are commonly used in foods to inhibit free radical damage to lipids. However, a concern has now been expressed about the safety of certain synthetic antioxidants such as BHA and BHT, which are suspected as possible carcinogens. As a result, BHA has been removed from the GRAS (Generally Recognized As Safe) list and the use of TBHQ remains prohibited in some countries. Therefore, there has been a considerable interest by the industry and a growing trend in consumer preferences for natural antioxidants over synthetic compounds (Madhavi, D. L.; Salunkhe, D. K. Toxicological Aspects of Food Antioxidants, In-Food Antioxidants, Eds.; Madavi, D. L., Deshpande, S. S. Salunkhe, D. K., Marcel Dekker Inc, New York, 1995, p.267). Therefore, the importance of search and exploitation of natural antioxidants, especially of plant origin has greatly increased in recent years (Jayaprakasha, G.K.; Jaganmohan Rao, L. Phenolic constituents from lichen Parmotrema stuppeum (Nyl.) Hale and their antioxidant activity. Zeitschrift fur Naturforschung 2000, 55c, 1018-1022). Dillenia a small genus of trees found in Indo - Malaysian region extending to Tropical Australia. About six species are found in India i.e. D. indica, D. aurea, D. pentagyna, D. andamanica, D. bractea and D. scabrella. Most of the species bear showy flowers and edible fruits and yield timbers of secondary importance (The Wealth of India, 1952). For the present investigation we have used the fruits of D. indica. D. indica occurs in the moist and evergreen forests of sub-Himalayan tract, from Kumaon and Garhwal eastwards to Assam, Bengal and Orissa. A handsome evergreen tree, 30-50 ft. in height and 6 ft. in girth, with a rounded crown. Leaf oblong-lanceolate, 8-14 in. long and 2-4 in. broad, with pointed apex and toothed margin; the upper part of the leaf as well as the veins beneath are covered with hairs. Flowers are large, 5-8 in. diameter, white, fragrant and solitary. Fruits are large 3-5 in. diameter, hard, consisting of five closely fitting imbricate sepals enclosing numerous seeds embedded in a gelatinous pulp (Figure 1 and 2). Seeds are small, compressed reniform with hairy margins. The tree flowers during May-August and fruits ripen during September-February. The sepals are sour and used in curries or made into jams and jellies. The acid juice is sweetened with sugar and used as cooling drink. The fruit is said to possess tonic and laxative properties and is used in abdominal pains. Literature survey revealed that, there is no report on the isolation of antioxidants from Dillenia indica. The present invention provides a process for preparation of an antioxidant fraction, which has antioxidant activity. Hence, we have developed a process for the preparation of antioxidants from the rinds of Dillenia indica. The principle of the present invention is to provide technology for the preparation of antioxidants from the rinds of Dillenia indica, which can be used as potential natural preservative and also supplemented in the food for general health benefits as a neutraceuticals / functional food. This invention is related to an efficient process for the preparation of antioxidants from the dried rinds of the fruits of Dillenia indica. The main object of the present invention is to provide a process for the preparation of antioxidant from Dillenia indica. Another object is to provide an efficient process which involves the use of simple extraction methods and solvents, which can be regenerated for the preparation of antioxidant from Dillenia indica. Accordingly the present invention provides a process for the preparation of antioxidants from Dillenia indica, which comprises: i) powdering the fruit rinds of Dillenia indica using mixer grinder to mesh size of 40-80, ii) extracting the above said material with hexane in a Soxhlet extractor at a temperature of 55-60 °C for a period of 6-10 h., iii) distilling the above said extract to recover up to 90% the solvent, iv) extracting the spent obtained from step (ii) with an alcoholic solvent in a Soxhlet extractor at a temperature of 55-60 °C for a period of 6-10 hrs., v) filtering the above said extract to obtain the particle free extract, vi) distilling the above said particle free extract at a temperature of 55 - 60°C to recover about 90% solvent, vii) drying the above concentrated extract using vacuum oven at a temperature of 40-50 °C, at a pressure of 10-25 mm of mercury under vacuum to obtain the desired brown colour product. In an embodiment of the present invention the alcoholic solvent used is selected from methanol and ethanol. i) In an another embodiment the Whatman filter paper No.l is used for the filtration of an extract in step v. In still another embodiment the yield of methanol extract was found to be 24-33%. The preparation of antioxidants from Dillenia indica was done according to following flow diagram Rinds of Dillenia indica Powdered using mixer grinder to get a mesh size of 40-80 Extraction with hexane using Soxhlet extractor for 6-10 h at 55-60 °C Hexane extract Spent I Distillation to recover hexane Extraction with Methanol (6-10 h at 55-60 °C) Methanol extract Spent Distillation at 55-60°C Concentrate Solvent Dried under vacuum at 40-50 °C Under 10-25 mm of mercury Yield of antioxidants (24-33%) The novelty of the process is 1. This is the first report of preparation of antioxidants from D. indica. 2. The invention is a one step process to obtain the antioxidants from D. indica. The advantages of the process is 1. The process is simple and the solvents used in this process can be regenerated for further use. The following examples are given by way of illustration of the present invention and therefore should not be constructed to limit the scope of the present invention. Example 1 50 g of rinds of Dillenia indica was powdered using mixer grinder to get a 60 mesh size. The powder was extracted with hexane for 6 h in a Soxhlet extractor at 60 °C. The extract was filtered and distilled to recover the solvent. The spent was dried and further extracted with 200 ml of methanol at 60 °C for 8 h in a Soxhlet extractor. The extract was filtered using Whatman filter paper No.l. The methanol extract was distilled under vacuum to recover the solvent. The extract was dried at a temperature of 50 °C and under reduced pressure at 10 mm of mercury. The yield of methanol extract was 15.05 (w/w). The phenolic content of the above extract was 34.2%. The extract obtained above method was screened for antioxidant activity using p-carotene-linoleate model system [Singh et al., J. Agricultural and Food Chemistry, 50, 81-86, 2002]. It showed 80.2% antioxidant activity at 100 ppm. Example -2 100 g of Dillenia indica was powdered using mixer grinder to get a 40 mesh size. The powder was extracted with hexane for 8 h in a Soxhlet extractor at 60 °C. The extract was filtered and distilled to recover the solvent. The spent was dried and further extracted with 400 ml of methanol at 60 °C for 6 h in a Soxhlet extractor. The extract was filtered using Whatman filter paper No.l. The methanol extract was distilled under vacuum to recover the solvent. The extract was dried at a temperature of 40 °C and under reduced pressure at 25 mm of mercury. The yield of methanol extract was 24.3 g. The phenolic content of the above extract was 27.34%. The extract obtained above method was screened for antioxidant activity using l,l-diphenyl-2-picrylhydrazyl (DPPH) [Singh et al., J. Agricultural and Food Chemistry, 50, 81-86, 2002]. It showed 68% antioxidant activity at 100 ppm using Diphenyl picrylhydrazyl (DPPH) method. Example -3 150 g of Dillenia indica was powdered using mixer grinder to get 80 mesh size. The powder was extracted with hexane for 10 h in a Soxhlet extractor at 55 °C. The extract was filtered and distilled to recover the solvent. The spent was dried and further extracted with 200 ml of methanol at 55 °C for 10 h in a Soxhlet extractor. The extract was filtered using Whatman filter paper No. 1. The methanol extract was distilled under vacuum to recover the solvent. The extract was dried at a temperature of 40 °C and under reduced pressure at 25 mm of mercury. The yield of methanol extract was 37.05 (w/w). The phenolic content of the above extract was 29.2%. The extract obtained above method was screened for antioxidant activity using l,l-diphenyl-2-picrylhydrazyl (DPPH) [Singh et al., J. Agricultural and Food Chemistry, 50, 81-86, 2002]. It showed 62.5% antioxidant activity at 50 ppm using Diphenyl picrylhydrazyl (DPPH) method. The antioxidant activities of the antioxidants from D. indica in comparison with BHA as been presented in Table 1 and 2. Table 1 Antioxidant activity of fruit extracts of D. indica and BHA by p-carotene-linoleate model system (% inhibition of bleaching of ß-carotene). (Table Removed) Table 2 Radical scavenging activity of fruit extracts of D. indica and BHA by DPPH method. (Table Removed) We Claim 1. A process for the preparation of from Dillenia indica, which comprises: i) powdering the fruit rinds of Dillenia indica using mixer grinder to mesh size of 40-80, ii) extracting the above said material with hexane in a Soxhlet extractor at a temperature of 55-60 °C for a period of 6-10 h., iii) distilling the above said extract to recover up to 90% the solvent, iv) extracting the spent obtained from step (ii) with an alcoholic solvent in a Soxhlet extractor at a temperature of 55-60 °C for a period of 6-10 hrs., v) filtering the above said extract to obtain the particle free extract, vi) distilling the above said particle free extract at a temperature of 55 - 60 °C to recover about 90% solvent, vii) drying the above concentrated extract using vacuum oven at a temperature of 40-50 °C, at a pressure of 10-25 mm of mercury under vacuum to obtain .the extract containg 24 to 33% antioxidants. 2. A process as claimed in claim 1, wherein the alcoholic solvent used is selected from methanol and ethanol. 3. A process as claimed in claims 1&2, wherein What man filter paper No.l is used for the filtration of an extract in step v. 4. A process for the preparation of antioxidants from Dillenia indica substantially as . herein described with reference to the examples.

Documents:

1091-del-2002-abstract.pdf

1091-del-2002-claims.pdf

1091-del-2002-correspondence-others.pdf

1091-del-2002-correspondence-po.pdf

1091-del-2002-description (complete).pdf

1091-del-2002-form-1.pdf

1091-del-2002-form-18.pdf

1091-del-2002-form-2.pdf

1091-del-2002-form-3.pdf

1091-del-2002-form-4.pdf

1091-del-2002-petition-137.pdf